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Morphologic investigation
Morphologic investigation together with fluorescence immunocytochemistry (ICC) is a common procedure for the identification and enumeration of CTCs after enrichment.
The CellSearch ® system classifies a CTC as positive event if the cell is ≥ 4 μm, DAPI+ (4,6-diamino-2-phenylindole), pan-keratin+, and CD45−.
The additional 4th fluorescence channel is accessible for a user-defined detection of, for example, therapy-relevant markers such as the androgen receptor (AR), (...)

Another form of non-EMT-associated dissemination, which is thought to substantially contribute to metastasis, is the release of "circulating tumor cell clusters" (Aceto et al, 2014).
These clusters are constituted of 2–50 tumor cells held together by plakoglobin-dependent intercellular adhesion.
Clustered cells are less likely to undergo anoikis and have an increased likelihood of being trapped in narrow blood vessels, thus favoring extravasation into distant (...)

Centrosome amplification has been linked to de-differentiation and recently been shown to induce invasion in cancer (Ghadimi et al, 2000; Lingle et al, 2002; Godinho et al, 2014).
The centrosomes form the main microtubule organizing center in mammalian cells and facilitate chromosomal separation during mitosis via the meiotic spindle apparatus but also participate in the organization of flagella and cilia.
EMT is thought not to play a role in the invasion of tumors with centrosome (...)

Metastasis, not the primary tumor, is responsible for the majority of breast cancer-related deaths. Emerging evidence indicates that breast cancer stem cells (CSCs) and the epithelial-to-mesenchymal transition (EMT) cooperate to produce circulating tumor cells (CTCs) that are highly competent for metastasis.
CTCs with both CSC and EMT characteristics have recently been identified in the bloodstream of patients with metastatic disease.
Breast CSCs have elevated tumorigenicity required for (...)